Identification and expression analysis of dmc1 in Chinese soft-shell turtle (Pelodiscus sinensis) during gametogenesis.

DNA meiotic recombinase 1 (disrupted meiotic cDNA, Dmc1) protein is homologous to the Escherichia coli RecA protein, was first identified in Saccharomyces cerevisiae. This gene has been well studied as an essential role in meiosis in many species. However, studies on the dmc1 gene in reptiles are limited. In this study, a cDNA fragment of 1,111 bp was obtained from the gonadal tissues of the Chinese soft-shell turtle via RT-PCR, containing a 60 bp 3' UTR, a 22 bp 5' UTR, and an ORF of 1,029 bp encoding 342 amino acids, named Psdmc1. Multiple sequence alignments showed that the deduced protein has high similarity (>95 %) to tetrapod Dmc1 proteins, while being slightly lower (86-88 %) to fish species.Phylogenetic tree analysis showed that PsDmc1 was clustered with the other turtles' Dmc1 and close to the reptiles', but far away from the teleost's. RT-PCR and RT-qPCR analyses showed that the Psdmc1 gene was specifically expressed in the gonads, and much higher in testis than the ovary, especially highest in one year-old testis. In situ hybridization results showed that the Psdmc1 was mainly expressed in the perinuclear cytoplasm of primary and secondary spermatocytes, weakly in spermatogonia of the testes. These results indicated that dmc1 would be majorly involved in the developing testis, and play an essential role in the germ cells' meiosis. The findings of this study will provide a basis for further investigations on the mechanisms behind the germ cells' development and differentiation in Chinese soft-shell turtles, even in the reptiles.

Associations between Ornithodoros spp. Ticks and Mojave Desert Tortoises (Gopherus agassizii) Obtained from Health Assessment Documents.

Soft ticks in the genus Ornithodoros occur throughout the Mojave Desert in southern Nevada, southeastern California, and parts of southwestern Utah and northwestern Arizona, USA, and are frequently observed parasitizing Mojave desert tortoises (Gopherus agassizii). However, limited research exists examining the relationship between ticks and desert tortoises. Mojave desert tortoises are listed as threatened by the US Fish and Wildlife Service, and as such, their populations are monitored and individual tortoise health is routinely assessed. These health assessments document the presence and abundance of ticks present on tortoises, but detailed examination of the relationship between ticks and tortoise health has been lacking. This study analyzed the relationship between tick presence and desert tortoise health assessments as a function of season, location, age (adult vs. juvenile), foraging behavior, evidence of clinical signs of disease, body condition score, and sex. Our results indicate that more ticks were found on tortoises in the summer than in any other season. Ticks were observed more frequently on captive tortoises versus wild tortoises, and more ticks were likely to be present on adult tortoises than on juveniles. Ticks were also more likely to be observed on tortoises that lacked evidence of foraging and on tortoises with observed clinical signs of disease. These findings provide valuable insights into the biology of ticks in relation to tortoises that may be useful for management of both captive and free-living threatened tortoise populations where ticks are detected. Our study also may improve understanding of potential tick-borne disease dynamics in the Mojave desert tortoise habitat, including Borrelia sp. carried by Ornithodoros ticks, which cause tick-borne relapsing fever in people.

Swimming through sand: using accelerometers to observe the cryptic, pre-emergence life-stage of sea turtle hatchlings.

Animals that hatch within a subterranean nest, such as turtle hatchlings, expend some of their limited energy reserves digging out through sand or soil to reach the surface. In sea turtles, this emergence process can take the hatchlings 3-7 days. However, we have a poor understanding of this process as it is difficult to observe what is occurring underground. Here, we utilize a novel method to characterize digging-out behaviour: affixing an accelerometer directly to newly hatched green turtles (Chelonia mydas) to record movement until nest emergence. Our data revealed that buried hatchlings maintained a head-up orientation but did not move in the expected left and right swaying motion associated with alternating limb crawling. Rather, they moved using dorsal-ventral heaving and pitching as if swimming vertically through the sand to the surface. Movement activity was irregular and brief, interspersed by many short periods of inactivity, mostly lasting less than 10 min. The first 24 h of head-up activity displayed no diel patterns, but the last 24 h prior to emergence involved more intense movement during night-time hours compared with daytime hours. Thus, our results add valuable new insight, and in some cases change previous assumptions, regarding the digging behaviours during the egg-to-emergence life stage in sea turtles.

Lung mechanics in juvenile and adult Chelonoidis carbonarius.

Testudines possess a rigid shell that influences the mechanics of the respiratory system. We studied respiratory mechanics in the terrestrial red-footed tortoise Chelonoidis carbonarius (Cryptodira), comparing juvenile individuals with a less ossified and more flexible carapace with adults with a well-ossified rigid shell. Combined with these ontogenetic differences, we analyzed respiratory system mechanics with animals in a supine and a prone position, as well as in the isolated lungs, to evaluate the impact of the viscera on breathing mechanics. To do so, we used established protocols to measure pulmonary volume (i.e. resting, VLr; and maximum, VLm), static (Cstat) and dynamic (Cdyn) compliance, and the work of breathing (W). We observed that isolated lungs displayed increased VLr, VLm, Cstat and Cdyn and decreased W. Additionally, pulmonary volume, compliance and W were affected by evaluated position, such as a smaller VLr in a supine position. Cdyn and W showed a volume dependency while frequency had less influence on these variables. At similar levels of ventilation, juveniles showed a lower W than adults when standardized by body mass, but similar W when standardized by VLr. Clear ontogenetic changes could be observed in breathing mechanics between juvenile and adult C. carbonarius. While these differences might largely be explained by variation in shell ossification, other explanations such as differences in visceral proportions or developmental degree of the post-pulmonary septum should also be taken into account.

Thecal and Epithecal Ossifications of the Turtle Shell: Ontogenetic And Phylogenetic Aspects.

The problem of the origin of the bony shell in turtles has a two-century history and still has not lost its relevance. First, this concerns the issues of the homology, the sources of formation and the ratio of bones of different nature, that is, thecal and epithecal, in particular. This article analyzes various views on the nature of the shell elements, and proposes their typification, based on modern data on developmental biology. It is proposed that the defining characteristic of the types of shell ossifications is not the level of their anlage in the dermis (thecality or epithecality), but, first of all, the primary sources of their formation: (1) neural crest (nuchal and plastral plates); (2) vertebral and rib periosteum (neural and costal plates); and (3) dermal mesenchyme (peripheral, suprapygal and pygal plates, as well as epithecal elements). In addition, there is complete correspondence between these types of ossifications and the sequence of their appearance in the turtle ontogenesis. The data show fundamental coincidence of the modifications of the ontogenetic development and evolutionary formation of the shell ossifications and are in agreement with a stepwise model for the origin of the turtle body plan. Particular attention is paid to the origin of the epithecal elements of the turtle shell, which correspond to the additional or supernumerary ossifications and seem to have wider distribution among turtles, than previously thought.

Epidemiological survey of gastrointestinal helminths and protozoa in Testudines from Sardinia, Italy.

The order Testudines comprises some of the most endangered groups of vertebrates. Under specific circumstances, infectious and parasitic diseases may affect the survival and fecundity of these animals, potentially threatening Testudines populations. In Sardinia, besides the three species of tortoises present in the wild (Testudo hermanni, Testudo graeca and Testudo marginata), many others are kept as pets. However, epidemiological studies on these animals have not been conducted so far. Thus, the aim of the work was to investigate the presence of gastrointestinal parasites in captive and wild tortoises of Sardinia, Italy, with particular regard to those of zoonotic importance. For the 215 animals examined (n = 36 wild caught and n = 179 private-owned), fecal samples were collected and processed by flotation and modified Ziehl-Neelsen technique. An overall prevalence of 81.4% for endoparasites was detected, with oxyurids being the most prevalent (74.4%), followed by Nyctotherus spp. (18.6%), Cryptosporidium spp. (12.6%), Angusticaecum spp. (2.8%), strongyles (0.9%), Balantidium spp. (0.9%), coccidia (0.9%), cestodes (0.5%), and Giardia spp. (0.5%). Data suggest that tortoises are affected by a great variety of endoparasites, and further molecular analysis are required to assess the impact of Cryptosporidium and Giardia species in these hosts. Therefore, regular health screenings are of importance for the management of these animals and for preventing emerging infectious diseases.

Overexpression of Kdm6b induces testicular differentiation in a temperature-dependent sex determination system.

In reptiles, such as the red-eared slider turtle ( Trachemys scripta elegans), gonadal sex determination is highly dependent on the environmental temperature during embryonic stages. This complex process, which leads to differentiation into either testes or ovaries, is governed by the finely tuned expression of upstream genes, notably the testis-promoting gene Dmrt1 and the ovary-promoting gene Foxl2. Recent studies have identified epigenetic regulation as a crucial factor in testis development, with the H3K27me3 demethylase KDM6B being essential for Dmrt1 expression in T. s. elegans. However, whether KDM6B alone can induce testicular differentiation remains unclear. In this study, we found that overexpression of Kdm6b in T. s. elegans embryos induced the male development pathway, accompanied by a rapid increase in the gonadal expression of Dmrt1 at 31°C, a temperature typically resulting in female development. Notably, this sex reversal could be entirely rescued by Dmrt1 knockdown. These findings demonstrate that Kdm6b is sufficient for commitment to the male pathway, underscoring its role as a critical epigenetic regulator in the sex determination of the red-eared slider turtle.

High thermal tolerance of egg clutches and potential adaptive capacity in green turtles.

Climate warming threatens sea turtles, among other effects, because high temperatures increase embryo mortality. However, not all species and populations are expected to respond the same way because they could have different thermal tolerances and capacities to adapt. We tested the effect of incubation temperature on egg mortality in a population of green turtles (Chelonia mydas) previously suggested to be less affected by extreme climatic events than others. We (1) assessed the relationship between temperature and hatching success, (2) defined an optimal range of temperatures that maximized hatching success and (3) assessed the variability in the response to temperature among clutches laid by different mothers, which could allow adaptation. Hatching success was consistently high in green turtle clutches with a skew toward high values, with 50 % of clutches having a success above 94 %. Yet, it was mildly affected by temperature, declining at both low and high temperatures. The optimal range of mean incubation temperatures was between ~30.5 °C and 32.5 °C. Current mean temperatures (31.3 °C) fall within the middle of the optimal range, indicating a potential resilience to further rises in mean nest temperature. Hatching success was best described by nest temperature and the interaction between female identity and temperature. This last predictor indicated a variability in thermal tolerance among clutches laid by different mothers and therefore, a capacity to adapt. The studied population of green turtles seems to be less vulnerable than others to climate warming. Understanding how different populations could respond to increasing temperatures could help complete the picture on the potential effects of climate change on sea turtles.

Shell biomechanics suggests an aquatic palaeoecology at the dawn of turtle evolution.

The turtle shell is a remarkable structure that has intrigued not only evolutionary biologists but also engineering and material scientists because of its multi-scale complexity and various functions. Although protection is its most apparent role, the carapace and plastron are also related to many physiological functions and their shape influences hydrodynamics and self-righting ability. As such, analysing the functional morphology of the shell could help understanding the ecology of Triassic stem-turtles, which will contribute to the century-long debate on the evolutionary origins of turtles. Here, we used 3D imaging techniques to digitize the shells of two of the earliest stem-turtle taxa, Proganochelys and Proterochersis, and submitted their models to biomechanical and shape analyses. We analysed the strength performance under five predation scenarios and tested the function of two morphological traits found in stem-turtles, the epiplastral processes and an attached pelvic girdle. The latter, also present in the crown-lineage of side-necked turtles, has been suggested to increase load-bearing capacity of the shell or to improve swimming in pleurodires. Our results do not confirm the shell-strengthening hypothesis and, together with the results of our shape analyses, suggest that at least one of the first stem-turtles (Proterochersis) was an aquatic animal.

Use of consumer insights to inform behavior change interventions aimed at illegal pet turtle trade in China.

Illegal poaching and overexploitation for the international pet trade are among the greatest threats to freshwater turtles in Southeast Asia. Expanding consumer research in China is crucial to filling knowledge gaps about the scale and structure of illegal trade and developing audience-targeted and relevant interventions that may reduce demand for illegal turtles as pets. We applied mixed methods to provide a detailed understanding of the consumer side of the illegal pet turtle trade in China. We conducted 30 interviews with key stakeholders and online surveys (n = 2456) of turtle keepers via community forums. From these, we identified 3 core consumer groups based on their prior turtle-keeping experience, species exposure, and potential for future purchases. We conducted a thematic qualitative analysis of concepts related to the capability, opportunity, motivation, and behavior (COM-B) model to determine the factors influencing the illegal pet turtle trade and to identify barriers to illegal purchases. Specifically, we identified purchasers' capabilities, opportunities, and motivations in the context of legality, enforcement risk, captive breeding, and impacts on wild population. We developed consumer journey maps (i.e., visual representations of customer's experiences throughout their buying journey) for core consumer groups. These maps illustrate the sequential behaviors and processes that consumers undertake when purchasing turtles, from initial exposure to sourcing, keeping, and providing a new home. Key factors influencing illegal purchases included convenient purchase channels, misguided cognition and motivations for pet keeping, and weak law enforcement. Effective interventions included messages focusing on shifting cognition and beliefs, increasing legal risk perception, and emphasizing stringent law enforcement, primarily delivered through online channels. Our results underscore the necessity for adaptable, audience-tailored interventions to reduce consumer demand for illegal wildlife products. The mixed-methods approach, combining quantitative and qualitative data, provided a comprehensive understanding of the target behavior and can inform the development of effective intervention strategies.

Uso de las percepciones del consumidor para guiar las intervenciones de cambio conductual enfocadas en el mercado ilegal de tortugas en China Resumen La caza furtiva y la sobreexplotación por el comercio internacional de mascotas son algunas de las mayores amenazas para las tortugas de agua dulce del sudeste asiático. Es esencial conocer más sobre los consumidores en China para llenar los vacíos en el conocimiento sobre la escala y la estructura del comercio ilegal y desarrollar intervenciones pertinentes y dirigidas al público que puedan reducir la demanda de tortugas como mascotas ilegales. Aplicamos métodos mixtos para conocer en detalle la percepción del consumidor del comercio ilegal de tortugas como mascotas en China. Realizamos 30 entrevistas a actores clave y encuestas en línea (n=2456) a cuidadores de tortugas a través de foros comunitarios. A partir de ellas, identificamos tres grupos principales de consumidores en función de su experiencia previa en la cría de tortugas, su exposición a las especies y su potencial para futuras compras. Realizamos un análisis cualitativo temático de conceptos relacionados con el modelo de capacidad, oportunidad, motivación y comportamiento (COM­B) para determinar los factores que influyen en el comercio ilegal de tortugas e identificar las barreras para las compras ilegales. En concreto, identificamos las capacidades, oportunidades y motivaciones de los compradores en el contexto de la legalidad, el riesgo de aplicación de la ley, la cría en cautiverio y el impacto sobre la población silvestre. Elaboramos mapas de viaje del consumidor (representaciones visuales de las experiencias del cliente a lo largo de su compra) para los principales grupos de consumidores. Estos mapas ilustran los comportamientos y procesos secuenciales que llevan a cabo los consumidores cuando compran tortugas, desde la exposición inicial hasta la adquisición, el mantenimiento y la asignación de un nuevo hogar. Entre los factores clave que influyen en las compras ilegales se encuentran la conveniencia de los canales de compra, los conocimientos y motivaciones erróneos para mantener mascotas y la debilidad de la aplicación de la ley. Las intervenciones eficaces incluyeron mensajes centrados en cambiar los conocimientos y las creencias, aumentar la percepción del riesgo legal y hacer hincapié en la aplicación rigurosa de la ley, principalmente a través de canales virtuales. Nuestros resultados destacan la necesidad de intervenciones adaptables y adaptadas al público para reducir la demanda de productos ilegales de fauna silvestre por parte de los consumidores. El enfoque de métodos mixtos, que combina datos cuantitativos y cualitativos, proporcionó una comprensión exhaustiva del comportamiento objetivo y puede servir de base para el desarrollo de estrategias de intervención eficaces.

Present and future distribution of the European pond turtle versus seven exotic freshwater turtles, with a focus on Eastern Europe.

Freshwater turtles are often used as terrarium pets, especially juveniles of exotic species. At the adult stage they are often released by their owners into the wild despite their high invasion potential. In Europe these thermophilic potentially invasive alien species occupy the habitats of the native European pond turtle Emys orbicularis (Linnaeus, 1758), with new records from the wild being made specifically in Eastern Europe (Latvia and Ukraine) during recent decades. Assessing the potential of alien freshwater turtles to establish in new territories is of great concern for preventing invasion risks while preserving native biodiversity in the present context of climate change. We explored this issue by identifying the present and future (by 2050) suitable habitats of the European pond turtle and several potentially invasive alien species of freshwater turtle already settled in Europe, using a geographic information system (GIS) modelling approach based on datasets from CliMond for climate, Near-global environmental information (NGEI) for freshwater ecosystems (EarthEnv) and Maxent modelling using open-access databases, data from the literature and original field data. Modelling was performed for seven species of alien freshwater turtles occurring from the extreme northern to southern borders of the European range of E. orbicularis: the pond slider Trachemys scripta (Thunberg and Schoepff, 1792), the river cooter Pseudemys concinna (Le Conte, 1830), the Florida red-bellied cooter Pseudemys nelsoni (Carr, 1938), the false map turtle Graptemys pseudogeographica (Gray, 1831), the Chinese softshell turtle Pelodiscus sinensis (Wiegmann, 1835), the Caspian turtle Mauremys caspica (Gmelin, 1774) and the Balkan terrapin Mauremys rivulata (Valenciennes, 1833). In Ukraine, the most Eastern limit of E. orbicularis distribution, were previously reported northern American originated T. scripta, M. rivulata, M. caspica, whereas in Latvia, Emys' most northern limit, were additionally reported P. concinna, P. nelsoni, G. pseudogeographica and Asia originated P. sinensis. The resulting Species Distribution Models (SDM) were of excellent performance (AUC > 0.8). Of these alien species, the most potentially successful in terms of range expansion throughout Europe were T. scripta (34.3% of potential range expansion), G. pseudogeographica (24.1%), and M. caspica (8.9%) and M. rivulata (4.3%) mainly in Eastern Europe, especially in the south of Ukraine (Odesa, Kherson, Zaporizhzhia regions, and Crimean Peninsula). Correlation between the built SDMs for the native E. orbicularis and the invasive alien T. scripta was reliably high, confirming the highly likely competition between these two species in places they cooccur. Moreover, a Multiple Regression Analysis revealed that by 2050, in most of Europe (from the western countries to Ukraine), the territory overlap between E. orbicularis and potentially invasive alien species of freshwater turtles will increase by 1.2 times, confirming higher competition in the future. Importantly, by 2050, Eastern Europe and Ukraine are predicted to be the areas with most suitable habitats for the European pond turtle yet with most limited overlap with the invasive alien species. We conclude that Eastern Europe and Ukraine are the most relevant priority conservation areas for the European pond turtle where it is now necessary to take protective measures to ensure safe habitat for this native species on the long-term.

AUTOLOGOUS OR ALLOGENIC BLOOD PLEURODESIS AS TREATMENT FOR PNEUMOCOELOM IN FOUR SEA TURTLES (LEPIDOCHELYS KEMPII, CARETTA CARETTA).

Pneumocoelom secondary to primary pulmonary disease was diagnosed via CT (three cases) or radiographs and coelioscopy (one case) in one Kemp's ridley (Lepidochelys kempii) and three loggerhead (Caretta caretta) sea turtles. All cases were suspected to be caused by trauma to the lung, and all exhibited positive buoyancy disorders. Coelomocentesis alone was ineffective at resolving each pneumocoelom, suggesting a large and persistent tear in the pulmonary parenchyma. An autologous (in three cases) or allogenic (in one case) blood patch pleurodesis was performed. The pneumocoelom was resolved in all four cases with no noted postoperative complications. Follow-up diagnostic imaging confirmed resolution of the pneumocoelom in three cases, and all four patients were successfully rehabilitated and had normal buoyancy at the time of release. One rehabilitated loggerhead was satellite tagged, and the tracking data demonstrated that the animal was still alive and moving normally 126 d postrelease, when the tag stopped transmitting. In sea turtles, an autologous or allogenic blood patch pleurodesis can be safely performed in cases of persistent pneumocoelom secondary to traumatic primary pulmonary disease.

Notes on the Courtship and Captive Reproduction of the Southern Vietnam Box Turtle, Cuora picturata, at the Turtle Conservation Center in Cuc Phuong National Park, Vietnam.

Endemic to only three known provinces in South Central Vietnam, Cuora picturata have faced extensive collection pressures for the food, medicinal, and pet trades. Further exacerbating their decline is the lack of protected areas where wild populations exist, with only one known population occurring within a protected area. With threats to wild populations persisting, the development of an assurance colony has been prioritized for C. picturata. The Asian Turtle Program of Indo-Myanmar Conservation has been operating the Turtle Conservation Center (TCC) in Cuc Phuong National Park since 1998. From 2020 to 2022, 24 C. picturata were acquired by the TCC from the illegal trade. With no ideal repatriation plan, these animals provided an opportunity for developing a captive assurance colony at the TCC to safeguard the species from extinction. As semi-aquatic species, box turtles in the genus Cuora are a unique group that shows variations in habitat use and behavior between species. Herein, we documented some specific courtship behaviors not yet described for C. picturata, providing insight into the unique reproductive behaviors of this species.

Moving Toward Evidence-Based Care: Measuring Preferences in Zoo-Housed Aquatic Turtles Using 24-h Remote Monitoring.

Providing evidence-based care to animals is a common goal in zoos and aquariums. However, there are many understudied species with a lack of information available, which may result in suboptimal care for individuals of these species. Using preference tests to ask individual animals what they want can be an effective means to enhance their care, and subsequently, their welfare. Using 24-h remote monitoring, we assessed substrate, enclosure feature, and lighting preferences of five aquatic turtles (two wood turtles, one Blanding's turtle, and two spotted turtles) that share an indoor enclosure at Lincoln Park Zoo. In addition, we compared enclosure feature and lighting preferences between winter and summer to determine whether the turtles had changing seasonal preferences despite being housed indoors. We found that the wood turtles and Blanding's turtle had substrate preferences, whereas the spotted turtles did not. In addition, we found that the turtles' enclosure feature preferences were consistent between winter and summer, but the amount of time spent on land, logs, and in water varied by individual. Finally, we found that the turtles utilized areas with ambient lighting more than areas with a heat lamp or UV lamp; this was consistent between winter and summer. The turtle preferences observed in this study can contribute to the development of care strategies that support their welfare and, in turn, may inspire similar preference studies at other institutions as we collectively advance toward providing animals of all taxonomic groups with evidence-based care.

Synovial fluid analysis in healthy green turtles Chelonia mydas in Taiwan.

Septic arthritis is a frustrating disease in sea turtle rehabilitation because of its unclear pathogenesis, delayed onset during rehabilitation, long-term treatment requirements, and potentially poor prognosis. Radiography, blood cultures, and arthrocentesis have been used as diagnostic tools for suspected cases. However, there is currently a lack of data on the characteristics of synovial fluid in healthy sea turtles. To establish reference data for synovial fluid in sea turtles, we enrolled 14 green turtles Chelonia mydas rescued between 2019 and 2022 from 3 facilities using the following inclusion criteria: normal attitude and appetite, normal motor functions of the 4 limbs, no joint swelling, and no ongoing use of antibiotics for at least 1 mo. Bacterial cultures of blood and synovial fluid from the shoulder joints of these turtles were obtained and a qualitative analysis of the synovial fluid was performed. The results revealed bacterial culture-negative blood and synovial fluids at 37°C. Most characteristics of normal synovial fluid in green turtles, such as being transparent, colorless, and able to create a strand of over 2.5 cm by being pulled with a needle in viscosity trials, as well as the cytology of the normal synovial fluids being dominated by histiocytes and synovial lining cells, lymphocytes, and occasionally a few heterophils or erythrocytes were similar to those in mammals. This study provides information on the normal synovial fluid characteristics of green turtles in Taiwan, which may be beneficial for the diagnosis of joint diseases in sea turtles.

Extensive sampling and citizen science expand the distribution of the threatened freshwater turtle Ranacephala hogei (Mertens, 1967).

Ranacephala hogei is a South American freshwater turtle considered one of the 25 most endangered chelonian species in the world. Endemic to the Atlantic Forest hotspot of southeastern Brazil, the conservation status of R. hogei is subject to continuous assessment at various levels. However, the scarcity of data regarding this species, particularly its geographic range, challenges these evaluations. In an effort to address these gaps, we conducted a comprehensive long-term inventory using different methods to study this species. Our efforts resulted in a 144% increase in documented occurrence points, including a new hydrographic basin and protected areas. By combining historical and current records, we have observed the persistence of R. hogei in rural areas, even in locations where its extinction was previously predicted. Consequently, our data significantly contribute to supporting future conservation assessments.

Toll-like Receptor Expression in Pelodiscus sinensis Reveals Differential Responses after Aeromonas hydrophila Infection.

Background: Toll-like receptor (TLR), as an important pattern recognition receptor, is a bridge between non-specific immunity and specific immunity, and plays a vital role in the disease resistance of aquatic animals. However, the function of TLR in Pelodiscus sinensis is still unclear. Methods and Results: The sequence characteristics and homology of three TLRs (PsTLR2, PsTLR3 and PsTLR5) were determined in this investigation. Their annotation and orthologies were supported by phylogenetic analysis, functional domain prediction, and sequence similarity analysis. qPCR showed that the identified TLRs were expressed in all tissues, among the high expression of PsTLR5 in the brain and liver and the high expression of PsTLR2 and PsTLR3 in the liver. PsTLR2 mRNA expression increased 6.7-fold in the liver 12 h after Aeromonas hydrophila infection, while the mRNA expression of PsTLR3 was down-regulated by 0.29 times in liver and 0.31 times in spleen. The mRNA expression of PsTLR5 was significantly up-regulated in four immune tissues, and it was up-regulated by 122.8 times in the spleen after 72 h infection. Finally, the recombinant proteins of extracellular LRR domains of these three TLRs were obtained by prokaryotic expression technology, and the binding tests were performed to discover their ability of binding pathogenic microorganisms. Microbial binding test showed that rPsTLR2, rPsTLR3 and rPsTLR5 can combine A. hydrophila, Edwardsiella tarda, Vibrio parahaemolyticus, Staphylococcus aureus, Streptococcus agalactiae and Candida albicans, while rPsTLR3 can bind A. hydrophila, E. tarda, V. parahaemolyticus and C. albicans. Conclusions: Our findings suggested that TLRs may be crucial to turtles' innate immune response against microbes.

Advancing Maternal Transfer of Organic Pollutants across Reptiles for Conservation and Risk Assessment Purposes.

Embryonic exposure through maternally transferred pollutants can affect embryo vitality, survival, and health. Reptiles face global declines and are sensitive to embryonic pollutant exposure. Yet, they are often neglected in pollution risk assessment and conservation. We analyzed maternal transfer of organic pollutants in reptiles through a systematic extraction, homogenization, and integration of published data on organic pollutants measured in mother-egg pairs into a comprehensive database (DOI:10.5281/zenodo.10900226), complemented with molecular physical-chemical properties of the pollutants. Over four decades, 17 publications provided 19,955 data points shifting from legacy to emerging contaminants although research on newer contaminants lags regulatory and societal demands. Challenges including taxonomic bias, heterogeneity in sampled tissues, and 73% of censored data complicate comparative analyses. However, significant opportunities were identified including the use of the turtle Malachlemys terrapin and snake Enhydris chinensis as flagship species where a large amount of data is available across tissues (allowing investigation into physiological relations) and compounds (allowing insights into maternal transfer across the chemical universe). Data on other freshwater and marine turtles provide the possibility of exploring taxonomic patterns in this subgroup. The analysis, integrated database, and discussion present opportunities for research in an era where science needs to achieve more with limited wildlife data.

Relationships between Spatial Biology and Physiological Ecology in the Gopher Tortoise, Gopherus polyphemus.

AbstractThe overlap between spatial and physiological ecology is generally understudied, yet both fields are fundamentally related in assessing how individuals balance limited resources. Herein, we quantified the relationships between spatial ecology using two parameters of home range (annual home range area and number of burrows used in 1 yr) and four measures of physiology that integrate stress and immunity (baseline plasma corticosterone [CORT] concentration, plasma lactate concentration, heterophil-to-lymphocyte [H∶L] ratio, and bactericidal ability [BA]) in a wild free-ranging population of the gopher tortoise (Gopherus polyphemus) to test the hypothesis that space usage is correlated with physiological state. We also used structural equation models (SEMs) to test for causative relationships between the spatial and physiological parameters. We predicted that larger home ranges would be negatively correlated with traditional biomarkers of stress and positively correlated with immunity, consistent with our hypothesis that home ranges are determined based on individual condition. Males had larger home ranges, used more burrows, and had higher baseline CORT than females. We found significant negative correlations between lactate and home range (r=-0.456, df=21, P=0.029). CORT was negatively correlated with the number of burrows used in both sexes (F=7.322, df=2,20, P=0.003, adjusted R2=0.383). No correlations were observed between space use and BA or, notably, H∶L ratio. SEMs suggested that variation in the number of burrows used was a result of variation in baseline CORT. The lack of a relationship between H∶L ratio and home range suggests that home range differences are not associated with differences in chronic stress, despite the pattern between baseline CORT and number of burrows used. Instead, this study indicates that animals balance trade-offs in energetics, likely by way of baseline corticosteroid, in such a way as to maintain function across continuously variable home range strategies.

pSTAT3 activation of Foxl2 initiates the female pathway underlying temperature-dependent sex determination.

Ovarian development was traditionally recognized as a "default" sexual outcome and therefore received much less scientific attention than testis development. In turtles with temperature-dependent sex determination (TSD), how the female pathway is initiated to induce ovary development remains unknown. In this study, we have found that phosphorylation of the signal transducer and activator of transcription 3 (pSTAT3) and Foxl2 exhibit temperature-dependent sexually dimorphic patterns and tempo-spatial coexpression in early embryos of the red-eared slider turtle (Trachemys scripta elegans). Inhibition of pSTAT3 at a female-producing temperature of 31 °C induces 64.7% female-to-male sex reversal, whereas activation of pSTAT3 at a male-producing temperature of 26 °C triggers 75.6% male-to-female sex reversal. In addition, pSTAT3 directly binds to the locus of the female sex-determining gene Foxl2 and promotes Foxl2 transcription. Overexpression or knockdown of Foxl2 can rescue the sex reversal induced by inhibition or activation of pSTAT3. This study has established a direct genetic link between warm temperature-induced STAT3 phosphorylation and female pathway initiation in a TSD system, highlighting the critical role of pSTAT3 in the cross talk between female and male pathways.